Effects of tricalcium silicate cements on osteogenic differentiation of human bone marrow-derived mesenchymal stem cells in vitro

Ashraf A. Eid, Khaled A. Hussein, Li Na Niu, Guo Hua Li, Ikuya Watanabe, Mohamed Al-Shabrawey, David Henry Pashley, Franklin Chi Meng Tay

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Tricalcium silicate cements have been successfully employed in the biomedical field as bioactive bone and dentin substitutes, with widely acclaimed osteoactive properties. This research analyzed the effects of different tricalcium silicate cement formulations on the temporal osteoactivity profile of human bone marrow-derived mesenchymal stem cells (hMW-MSCs). These cells were exposed to four commercially available tricalcium silicate cement formulations in osteogenic differentiation medium. After 1, 3, 7 and 10 days, quantitative real-time polymerase chain reaction and Western blotting were performed to detect expression of the target osteogenic markers ALP, RUNX2, OSX, OPN, MSX2 and OCN. After 3, 7, 14 and 21 days, alkaline phosphatase assay was performed to detect changes in intracellular enzyme level. An Alizarin Red S assay was performed after 28 days to detect extracellular matrix mineralization. In the presence of tricalcium silicate cements, target osteogenic markers were downregulated at the mRNA and protein levels at all time points. Intracellular alkaline phosphatase enzyme levels and extracellular mineralization of the experimental groups were not significantly different from the untreated control. Quantitative polymerase chain reaction results showed increases in downregulation of RUNX2, OSX, MSX2 and OCN with increasing time of exposure to the tricalcium silicate cements, while ALP showed peak downregulation at day 7. For Western blotting, OSX, OPN, MSX2 and OCN showed increased downregulation with increased exposure time to the tested cements. Alkaline phosphatase enzyme levels generally declined after day 7. Based on these results, it is concluded that tricalcium silicate cements do not induce osteogenic differentiation of hBM-MSCs in vitro.

Original languageEnglish (US)
Pages (from-to)3327-3334
Number of pages8
JournalActa biomaterialia
Volume10
Issue number7
DOIs
StatePublished - Jan 1 2014

Fingerprint

Silicate Cement
Bone cement
Stem cells
Mesenchymal Stromal Cells
Silicates
Bone
Cements
Bone Marrow
Down-Regulation
Phosphatases
Alkaline Phosphatase
Polymerase chain reaction
Enzymes
Assays
Western Blotting
Bone Substitutes
Alizarin
Dentin
Extracellular Matrix
tricalcium silicate

Keywords

  • Bone marrow
  • Human origin
  • Mesenchymal stem cells
  • Osteogenic differentiation
  • Tricalcium silicate cements

ASJC Scopus subject areas

  • Biotechnology
  • Biomaterials
  • Biochemistry
  • Biomedical Engineering
  • Molecular Biology

Cite this

Effects of tricalcium silicate cements on osteogenic differentiation of human bone marrow-derived mesenchymal stem cells in vitro. / Eid, Ashraf A.; Hussein, Khaled A.; Niu, Li Na; Li, Guo Hua; Watanabe, Ikuya; Al-Shabrawey, Mohamed; Pashley, David Henry; Tay, Franklin Chi Meng.

In: Acta biomaterialia, Vol. 10, No. 7, 01.01.2014, p. 3327-3334.

Research output: Contribution to journalArticle

Eid, Ashraf A. ; Hussein, Khaled A. ; Niu, Li Na ; Li, Guo Hua ; Watanabe, Ikuya ; Al-Shabrawey, Mohamed ; Pashley, David Henry ; Tay, Franklin Chi Meng. / Effects of tricalcium silicate cements on osteogenic differentiation of human bone marrow-derived mesenchymal stem cells in vitro. In: Acta biomaterialia. 2014 ; Vol. 10, No. 7. pp. 3327-3334.
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AU - Li, Guo Hua

AU - Watanabe, Ikuya

AU - Al-Shabrawey, Mohamed

AU - Pashley, David Henry

AU - Tay, Franklin Chi Meng

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AB - Tricalcium silicate cements have been successfully employed in the biomedical field as bioactive bone and dentin substitutes, with widely acclaimed osteoactive properties. This research analyzed the effects of different tricalcium silicate cement formulations on the temporal osteoactivity profile of human bone marrow-derived mesenchymal stem cells (hMW-MSCs). These cells were exposed to four commercially available tricalcium silicate cement formulations in osteogenic differentiation medium. After 1, 3, 7 and 10 days, quantitative real-time polymerase chain reaction and Western blotting were performed to detect expression of the target osteogenic markers ALP, RUNX2, OSX, OPN, MSX2 and OCN. After 3, 7, 14 and 21 days, alkaline phosphatase assay was performed to detect changes in intracellular enzyme level. An Alizarin Red S assay was performed after 28 days to detect extracellular matrix mineralization. In the presence of tricalcium silicate cements, target osteogenic markers were downregulated at the mRNA and protein levels at all time points. Intracellular alkaline phosphatase enzyme levels and extracellular mineralization of the experimental groups were not significantly different from the untreated control. Quantitative polymerase chain reaction results showed increases in downregulation of RUNX2, OSX, MSX2 and OCN with increasing time of exposure to the tricalcium silicate cements, while ALP showed peak downregulation at day 7. For Western blotting, OSX, OPN, MSX2 and OCN showed increased downregulation with increased exposure time to the tested cements. Alkaline phosphatase enzyme levels generally declined after day 7. Based on these results, it is concluded that tricalcium silicate cements do not induce osteogenic differentiation of hBM-MSCs in vitro.

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